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Effects of TGF-ß2 on Immune Response–Related Gene Expression Profiles in the Human Corneal Endothelium

¹From the Department of Corneal Tissue Regeneration and the ²Department of Ophthalmology, Tokyo University Graduate School of Medicine, Tokyo, Japan.

PURPOSE. To determine the effects of transforming growth factor (TGF)-ß2 on immune-response–related gene expression profiles in the stimulated human corneal endothelium (HCE).

METHODS. A human complementary DNA (cDNA) expression array analysis was used to investigate the effects of TGF-ß2 on cultured HCE incubated with interleukin (IL)-1 and tumor necrosis factor (TNF)-. Gene-specific semiquantitative reverse transcription–polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to examine the gene expression patterns revealed by the cDNA expression array analysis. Moreover, the expression of newly identified genes in HCE was confirmed by RT-PCR in human donor corneas.

RESULTS. cDNA expression array analysis and semiquantitative RT-PCR revealed that TGF-ß2 downregulated the expression of IL-6, growth-related (Gro)- (CXCL1), monocyte chemotactic protein (MCP)-1 (CCL2), granulocyte-colony stimulating factor (G-CSF), and insulin-like growth factor binding protein (IGFBP)-5 and upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-1. ELISA confirmed TGF-ß2–mediated changes in the expression of IL-6, CXCL1, CCL2, G-CSF, IGFBP-5, and TIMP-1 at the protein level. CXCL1, G-CSF, and IGFBP-5 mRNAs were detected for the first time in the HCE of donor corneas.

CONCLUSIONS. TGF-ß2 downregulates IL-6, CXCL1, CCL2, G-CSF, and IGFBP-5, and upregulates TIMP-1 in cultured HCE stimulated with proinflammatory cytokines, suggesting the immunomodulatory role of TGF-ß2 in the aqueous humor and the pathophysiological significance of TGF-ß2 in the anterior chamber of the eye.

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